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Leica tauSTED super resolution microscopy for molecular imaging in fixed and live specimens

Leica tauSTED 超分辨率显微镜，用于固定样本和活体样本的分子成像

基本信息

批准号：
10425103
负责人：
AMMASI PERIASAMY
金额：
$ 85.04万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-15 至 2023-06-14
项目状态：
已结题

项目摘要

Project Summary Super-Resolution Fluorescence Microscopy (SRFM) has become an indispensable tool for scientists to probe discrete molecular proximities. Currently, systems in the Keck Center at the University of Virginia can only reach resolutions of ≥120 nm; to break that barrier for fixed or live cells, Leica STELLARIS 8 tauSTED microscopy can achieve ~30-50 nm resolution for fixed specimens. As a research and teaching institution in the highly competitive academic environment, it is essential to make these advanced technologies available for the biomedical sciences. A second driver for this application is the need to replace the existing and non-repairable 14-year-old Leica SP5x confocal microscopy system, purchased through the S10 program in 2007. The requested super-resolution tauSTED microscopy system will answer both needs and will benefit the basic and clinical sciences of this university; in particular, the 12 participating federally funded investigators (9 major and 3 minor users) of the Keck Center (KCCI). The higher quantum efficiency HyD (46%) and the new photon detection efficiency (PDE) detector (58%) provides a better option to excite the sample with less excitation intensity and detecting weaker photon levels in cells and tissues. This instrument is essential for our core investigator group to investigate the subcellular architecture and cell dynamics at nanoscale dimensions and will also be critical for training other investigators and students attending the fall semester microscopy course interested in the study of various biological applications including developmental, neuroscience and cell biology. The requested system will be housed in the internationally recognized W.M. Keck Center for Cellular Imaging (KCCI), which is particularly known for its advanced microscopy and molecular imaging. KCCI is a university-wide imaging facility, located within walking distance of all participating investigators from the School of Arts & Sciences, Engineering, and Medicine. KCCI is also recognized for its annual hands-on training FLIM & FRET workshop, held for the past 19 years at the University of Virginia. The requested super resolution tauSTED microscopy is an advanced technology, which will be housed in the Leica SP5x room (11'x14') and configured appropriately to meet the experimental conditions of users in this proposal and additional existing and future users of the KCCI imaging core. In summary, the requested instrument will provide the research community at the University of Virginia and beyond with a level of imaging capability that is not currently available and will enhance significantly ongoing and future research.

项目摘要超分辨率荧光显微镜（SRFM）已成为一种不可或缺的工具，科学家来探测离散的分子邻近性。目前，在凯克中心的系统，弗吉尼亚大学只能达到≥120 nm的分辨率;为了打破固定或活细胞的这一障碍， Leica STELLARIS 8 tauSTED显微镜可为固定样本实现约30-50 nm的分辨率。作为在竞争激烈的学术环境中，研究和教学机构，这些先进的技术可用于生物医学科学。此应用程序的第二个驱动程序是需要更换现有的和不可修复的14岁的徕卡SP 5x共聚焦显微镜 2007年通过S10计划购买的系统。所要求的超分辨率tauSTED 显微镜系统将回答这两个需求，并将有利于基础和临床科学，这大学;特别是12名联邦资助的参与调查员（9名主要用户和3名次要用户）凯克中心（KCCI）更高的量子效率HyD（46%）和新的光子检测效率（PDE）检测器（58%）提供了更好的选择，以较少的激发激发样品强度和检测细胞和组织中较弱的光子水平。这一工具对我们的核心研究小组，研究纳米尺度下的亚细胞结构和细胞动力学维度，对于培训其他调查人员和参加秋季课程的学生也至关重要学期显微镜课程感兴趣的各种生物应用的研究，包括发育、神经科学和细胞生物学。所要求的系统将安装在国际公认的W. M.凯克中心细胞成像（KCCI），特别是其先进的显微镜和分子显像KCCI是一个大学范围内的成像设施，位于步行距离内的所有参与来自文理学院、工程学院和医学院的调查人员。KCCI也被公认为其年度实践培训FLIM & FRET研讨会，在过去的19年里在大学举行，弗吉尼亚所要求的超分辨率tauSTED显微镜是一种先进的技术，将是放置在Leica SP 5x室（11“x14”）中，并进行适当配置，以满足实验要求本提案中的用户以及KCCI成像核心的其他现有和未来用户的条件。总之，所要求的工具将为哥伦比亚大学的研究界提供弗吉尼亚州和超越的成像能力水平，目前还没有，将提高正在进行的和未来的研究。